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中性蛋白酶分步酶解花生分离蛋白制备花生短肽的研究
引用本文:李宁,刘红芝,刘丽,王强. 中性蛋白酶分步酶解花生分离蛋白制备花生短肽的研究[J]. 中国农业科学, 2013, 46(24): 5237-5247. DOI: 10.3864/j.issn.0578-1752.2013.24.018
作者姓名:李宁  刘红芝  刘丽  王强
基金项目:农业部公益性行业科研专项(201203037)、国家国际科技合作专项项目(2012DFA31400)
摘    要:【目的】为了充分利用花生榨油之后的副产物,提高产品附加值,建立花生短肽制备工艺,研发功能性花生短肽产品。【方法】通过比较酶种类、底物浓度、酶解温度、酶解时间对水解度与短肽得率的影响,采用二次回归正交旋转组合设计优化分步酶解制备花生短肽的最佳工艺。【结果】中性蛋白酶分步酶解花生分离蛋白制备短肽的最佳工艺参数为:Neutrase水解花生分离蛋白2.04 h后加入Protamex继续酶解1.96 h,Neutrase添加量为5 200 U•g-1底物,Protamex添加量为422.32 U•g-1 底物,水解温度44.83℃,底物浓度8%,在此条件下,短肽得率为83.93%,水解度为38.25%,花生短肽纯度为93.85%±0.44%。经高效液相色谱测定,分子量小于1 000 D的水解产物占98.88%。【结论】采用Neutrase与Protamex分步酶解花生分离蛋白制备花生短肽,与现有碱性蛋白酶酶解制备花生短肽方法相比,避免了后续脱盐步骤,简化了工艺,且具有制备条件温和,DH和TCA-NSI高,纯度高,分子量集中分布于1 000 D以下等特点。

关 键 词:中性蛋白酶   花生分离蛋白   分步酶解   花生短肽
收稿时间:2013-04-10

Preparation of Peanut Oligopeptides from Peanut Protein Isolated by Neutral Proteinase Stepping Hydrolysis
LI Ning,LIU Hong-Zhi,LIU Li,WANG Qiang. Preparation of Peanut Oligopeptides from Peanut Protein Isolated by Neutral Proteinase Stepping Hydrolysis[J]. Scientia Agricultura Sinica, 2013, 46(24): 5237-5247. DOI: 10.3864/j.issn.0578-1752.2013.24.018
Authors:LI Ning  LIU Hong-Zhi  LIU Li  WANG Qiang
Affiliation:Institute of Agro-Food Science and Technology, Chinese Academy of Agricultural Sciences/ Key Laboratory of Agricultural Product Processing and Quality Control, Ministry of Agriculture, Beijing 100193
Abstract:【Objective】The objective of this study is to make full use of the by-products after commercial extraction of peanut oil, and to increase the product additive value, as well as to establish the productive technology of peanut oligopeptides, and finally to put peanut oligopeptides in product of research and development. 【Method】The kind of enzyme, substrate concentration, enzymolysis temperature, enzymatic hydrolysis time on the influence of DH and TCA-NSIwere compared and the best preparation technology was determined by quadratic regression orthogonal rotational combing design. 【Result】 The optimum process parameters for the preparation of peanut oligopeptides from peanut protein isolated by neutral proteinase stepping hydrolysis was that Protamex hydrolyzed continually for 120 min after Neutrase hydrolyzed peanut protein isolated 120 min, the volume of addition of Neutrase was 5 200 U•g-1 substrate and the volume of addition of Protamex was 422.32 U•g-1 substrate, the enzymolysis temperature was 44.83℃, the substrate concentration was 8%. Under these conditions, the TCA-NSI was 83.93%, the DH was 38.25%, and the peanut oligopeptides purity was 93.85%±0.44%. Hydrolysis products with the relative molecular mass less than 1 000 D accounted for 98.88% by HPLC. 【Conclusion】Compared with the existing alkaline protease enzymatic preparation of peanut oligopeptides, the preparation of peanut oligopeptides from peanut protein isolated by Neutrase and Protamex stepping hydrolysis had the characteristics of avoiding subsequent desalting, simplifying process, preparation moderate conditions, high DH, TCA-NSI and purity, and the molecular weight of peptides being mainly concentrated at below 1 000 D.
Keywords:neutral proteinase    peanut protein isolated    stepping hydrolysis    peanut oligopeptides
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